Project Summary This shared instrument grant is to request funds for an BD Symphony that will support research at Yale University School of Medicine. There is a growing demand for high-parameter flow cytometry at Yale to characterize biological samples as completely as possible. A major goal of the NIH is to increase the amount of translational research being conducted in cancer immunology and tumor microenvironments, and consequently, the study of immune responses to cancer is increasing at Yale, as is research of other complex disease processes. As the amount of published phenotypic information on distinct cell types and their subsets continues to increase, the number of parameters that is then needed to uniquely identify and distinguish diverse cell subset is likewise growing at a rapid pace. Although many investigators now require a large amount of information about the cellular compositions to draw strong conclusions in their studies, the amount of tissue collected and the number of cells within the sample can be small. As a result, the goal is to obtain more phenotypic information in a single stain set that for many studies must be analyzed by fluorescence flow cytometry. To meet the increased demand for more fluorophores, new fluorescent dyes are being released that can be excited by new laser lines, which together will greatly expanded the number of parameters that can be detect by flow cytometry. Currently Yale does not have the capability to enable traditional flow cytometry with the number of parameters sought by many NIH funded investigators. There is strong institutional support for this equipment and the BD Symphony is an ideal instrument to meet these needs because it 1) can house up to 10 lasers, allowing us to install additional lasers in the future as new dyes emerge, 2) can accommodate up to 10 detectors off any laser line, with a maximum of 50 detectors, 3) is highly customizable and easily upgraded, and 4) enables state-of-the-art high- parameter flow cytometry to measure at least 20 cellular parameters simultaneously with the 6 lasers requested. Lastly, the vendor BD uniquely offers 25 options for laser lines that are not currently available from any other vendor of flow cytometers. Thus, the requested equipment for higher parameter fluorescence analysis will allow Yale researchers to interrogate complex biology with deeper phenotypic data sets not otherwise obtainable, accelerating the pace of research while engendering greater insights and deeper knowledge.